Mineral trioxide aggregate stimulates macrophages and mast cells to release neutrophil chemotactic factors: role of IL-1 beta, MIP-2 and LTB4

Objective. In the present study, the role of macrophages and mast cells in mineral trioxide aggregate (MTA)-induced release of neutrophil chemotactic factor was investigated.Study design. MTA suspension (50 mg/mL) was plated over inserts on macrophages or mast cells for 90 minutes. Untreated cells served as controls. Cells were washed and cultured for 90 minutes in RPMI without the stimuli. Macrophages and mast cell supernatants were injected intraperitoneally (0.5 mL/cavity), and neutrophil migration was assessed 6 hours later. In some experiments, cells were incubated for 30 minutes with dexamethasone (DEX, 10 mu M/well), BWA4C (BW, 100 mu M/well) or U75302 (U75, 10 mu M/well). The concentration of Leukotriene B-4 (LTB4) in the cell-free supernatant from mast cells and macrophage culture was measured by ELISA.Results. Supernatants from MTA-stimulated macrophages and mast cells caused neutrophil migration. The release of neutrophil chemotactic factor by macrophages and mast cells was significantly inhibited by DEX, BW, or U75. Macrophages and mast cells expressed mRNA for interleukin-1 (IL-1)beta and macrophage inflammatory protein-2 (MIP-2) and the pretreatment of macrophages and mast cells with DEX, BW, or U75 significantly altered IL-1 beta and MIP-2 mRNA expression. LTB4 was detected in the MTA-stimulated macrophage supernatant but not mast cells.Conclusions. MTA-induces the release of neutrophil chemotactic factor substances from macrophages and mast cells with participation of IL-1 beta, MIP-2, and LTB4. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: e135-e142)

Antigenic Activity of Bacterial Endodontic Contents from Primary Root Canal Infection with Periapical Lesions against Macrophage in the Release of Interleukin-1 beta and Tumor Necrosis Factor alpha

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Participation of alpha-1 and alpha-2 adrenoceptors of the lateral hypothalamic area in water intake, and renal sodium, potassium and urinary Volume excretion induced by central administration of angiotensin II

The circumventricular structures and the lateral hypothalamus (LH) have been shown to be important for the central action of angiotensin II (ANGII) on water and electrolyte regulation. Several anatomical findings have demonstrated neural connection between circumventricular structures and the LH, the present experiments were conducted to investigate the role of the alpha-adrenergic antagonists and agonistic injected into the LH on the water intake, sodium and potassium excretion elicited by injections of ANGII into the lateral ventricle (LV), the water intake was measured every 30 min over a period of 120 min. The sodium, potassium and urinary volume were measured over a period of 120 min in water-loaded rats. The injection of ANGII into the LV increased the water intake, which was reduced by previous injection of clonidine (an alpha-2-adrenergic agonist) into the LH. The injection of yohimbine (an alpha-2-adrenergic antagonist) and prazosin (an alpha-l-adrenergic antagonist) into the LH, which was done before injecting ANGII into the LV, also reduced the water intake induced by ANGII. The injection of ANGII into the LV reduced the sodium, potassium and urinary volume. Previous treatment with clonidine attenuated the action of ANGII in reducing the sodium, potassium and urinary volume, whereas previous treatment with yohimbine attenuated the effects of ANGII but with less intensity than that caused by clonidine. Previous treatment with prazosin increased the inhibitory effects of ANGII in those parameters. The injection of yohimbine and prazosin, which was done before the injection of clonidine, attenuated the effect of clonidine on the ANGII mechanism. The results of this study led us to postulate that when alpha-2-adrenergic receptors are blocked, the clonidine may act on the imidazoline receptors to produce its effects on the ANGII mechanism. We may also conclude that the LH is involved with circumventricular structures, which present excitatory and inhibitory mechanisms. Such mechanisms are responsible for regulating the renal excretion of sodium, potassium and water, (C) 2000 Elsevier B.V.

Functional characterisation of alpha(1)-adrenoceptors in denervated rat vas deferens

We investigated whether or not surgical denervation of the rat vas deferens changes the alpha(1)-adrenoceptor subtypes involved in the contractions to noradrenaline. Denervated vas deferens was approximate to22 times more sensitive to noradrenaline (pD(2)=7.35 +/- 0.04) than control vas (pD(2)= 6.01 +/- 0.03). This difference in noradrenaline potency was eliminated when cocaine (6 muM) was added to control vas (pD(2)=7.22 +/- 0.04). The noradrenaline-induced contractions of control and denervated vas deferens were insensitive to the alpha(1B)/alpha(1D)-adrenoceptor alkylating agent chloroethylclonidine (100 muM, 45 min). The concentration-response curves to noradrenaline in control and denervated vas deferens were competitively antagonised by prazosin (pA(2)approximate to9.6), WB-4101 (pA(2)approximate to9.5), 5-methyl urapidil (pA(2)approximate to8.4), phentolamine (pA(2)approximate to8.7), yohimbine (pA(2)approximate to6.9), BMY 7378 (pA(2)approximate to6.9) and indoramin (pA(2)approximate to8.7). After the treatment of control and denervated vas deferens with phenoxybenzamine, the partial agonist oxymetazoline antagonised competitively the concentration-response curves to noradrenaline showing pA(2) values approximate to7.4 in both groups. We conclude that noradrenaline-induced contractions in control and denervated rat vas deferens are mediated by alpha(1A)-adrenoceptors and that surgical denervation of the rat vas deferens is not able to change the alpha(1)-adrenoceptor subtypes involved in the contractions to noradrenaline.

Evolutionary and biogeographic history of the subfamily Neoplecostominae (Siluriformes: Loricariidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Is the evolutionary history of the O-type P element in the saltans and willistoni groups of Drosophila similar to that of the canonical P element?

We studied the occurrence of O-type P elements in at least one species of each subgroup of the saltans group, in order to better understand the phylogenetic relationships among the elements within the saltans group and with those of species belonging to the willistoni group. We found that the O-type subfamily has a patchy distribution within the saltans group (it does not occur in D. neocordata and D. emarginata), low sequence divergence among species of the saltans group as well as in relation to species of the willistoni group, a lower rate of synonymous substitution for coding sequences compared to Adh, and phylogenetic incongruities. These findings suggest that the evolutionary history of the O-type subfamily within the saltans and willistoni groups follows the same model proposed for the canonical subfamily of P elements, i.e., events of horizontal transfer between species of the saltans and willistoni groups.

Time-dependent expression of annexin 1 in a model of chronic granulomatous inflammation

Objective and design: To determine the expression pattern and distribution of the glucocorticoid-inducible protein annexin 1 (ANXA1) in a murine model of chronic granulomatous inflammation.Materials or subjects: TO Mouse.Treatment: Chronic granulomatous inflammation was induced by injecting into dorsal sub-cutaneous air-pouches in mice, a mixture of croton oil and Freund's complete adjuvant (CO/FCA).Methods: Western and northern analysis, corticosterone assay, and immunohistochemistry. Statistical analysis was performed using ANOVA followed by Tukey's pair-wise comparisons or Dunnett's multiple comparisons.Results: ANXA1 protein levels changed significantly throughout the 4-week time course, with an initial peak at day 7 and a later elevation at 28 days. ANXA1 mRNA levels peaked at days 1 and 3, with a significant decline at day 7 followed by an upward trend to day 28. Plasma corticosterone measurements taken throughout the time course revealed an increase from 14 days onward, suggesting that corticosterone does not influence ANXA1 expression during the initial stages of the model. Immunogold staining revealed that ANXA1 expression in the inflamed tissue was mainly in extravasated neutrophils, with intact protein (37 kDa) being predominantly observed on the cell membrane.Conclusions: the pattern of ANXA1 expression indicates that infiltrated neutrophils are responsible for the majority of ANXA1 present both at early and later stages of this model of granulomatous inflammation.

Properties of l=1 B-1 and B*(2) Mesons

This Letter presents the first strong evidence for the resolution of the excited B mesons B-1 and B-2(*) as two separate states in fully reconstructed decays to B+(*())pi(-). The mass of B-1 is measured to be 5720.6 +/- 2.4 +/- 1.4 MeV/c(2) and the mass difference Delta M between B-2* and B-1 is 26.2 +/- 3.1 +/- 0: 9 MeV/c(2), giving the mass of the B-2* as 5746.8 +/- 2.4 +/- 1.7 MeV/c(2). The production rate for B-1 and B-2* mesons is determined to be a fraction (13.9 +/- 1.9 +/- 3.2)% of the production rate of the B+ meson.

Nutritionally induced changes in endosperm of shrunken-1 and brittle-2 maize kernels grown in vitro

Although mineral nutrition affects maize (Zea mays L.) yield by controlling starch deposition in kernels, the mechanisms involved are largely unknown. Our objectives were to examine this relationship by nutritionally and genetically altering starch production in the endosperm. Kernels of W64A and two starch-deficient mutants, shrunken-1 and brittle-2, were grown in vitro with varying supplies of N (0-50 mM) or P (0-6 mM) to produce different degrees of endosperm starch production, and the levels of enzyme activities and metabolites associated with carbohydrate and N metabolism were examined. In vitro grown kernels exhibited the expected starch phenotypes, and a minimum level of media N (25 mM) and P (2 mM) was required for optimal growth. However, increasing the availability of N or P could not overcome the genetically induced decrease in starch deposition of the mutants. Nitrogen deficiency enhanced sugar accumulation, but decreased amino acid levels, soluble protein, enzyme activity, starch synthesis, and endosperm dry weight. Phosphorous deficiency also decreased starch production and endosperm dry weight, but with only a minimal effect on the activities of ADP-glucose pyrophosphorylase and alanine transaminase. Genotypic differences in endosperm starch, and the increases induced by N and P supply, Here closely associated with the level of endosperm N, but not endosperm P. Thus, while both N and P are crucial for optimal yield of maize grain, they appear to act by different means, and with different importance in governing starch deposition in the endosperm.

Quantization of (2+1)-spinning particles and bifermionic constraint problem

This work is a natural continuation of our recent study in quantizing relativistic particles. There it was demonstrated that, by applying a consistent quantization scheme to the classical model of a spinless relativistic particle as well as to the Berezin-Marinov model of a 3 + 1 Dirac particle, it is possible to obtain a consistent relativistic quantum mechanics of such particles. In the present paper, we apply a similar approach to the problem of quantizing the massive 2 + 1 Dirac particle. However, we stress that such a problem differs in a nontrivial way from the one in 3 + 1 dimensions. The point is that in 2 + 1 dimensions each spin polarization describes different fermion species. Technically this fact manifests itself through the presence of a bifermionic constant and of a bifermionic first-class constraint. In particular, this constraint does not admit a conjugate gauge condition at the classical level. The quantization problem in 2 + 1 dimensions is also interesting from the physical viewpoint (e.g., anyons). In order to quantize the model, we first derive a classical formulation in an effective phase space, restricted by constraints and gauges. Then the condition of preservation of the classical symmetries allows us to realize the operator algebra in an unambiguous way and construct an appropriate Hilbert space. The physical sector of the constructed quantum mechanics contains spin-1/2 particles and antiparticles without an infinite number of negative-energy levels, and exactly reproduces the one-particle sector of the 2 + 1 quantum theory of a spinor field.

Molecular models of protein kinase 6 from Plasmodium falciparum

Cyclin-dependent kinases (CDKs) have been identified as potential targets for development of drugs, mainly against cancer. These studies generated a vast library of chemical inhibitors of CDKs, and some of these molecules can also inhibit kinases identified in the Plasmodium falciparum genome. Here we describe structural models for Protein Kinase 6 from P. falciparum (PfPK6) complexed with Roscovitine and Olomoucine. These models show clear structural evidence for differences observed in the inhibition, and may help designing inhibitors for PfPK6 generating new potential drugs against malaria.

Subclinical abnormalities of glucose metabolism in Brazilian women with a history of gestational diabetes mellitus

Background. To evaluate insulin release and insulin sensitivity in women with prior gestational diabetes mellitus (GDM) to gain a better understanding of type 2 diabetes pathogenesis.Methods. GDM women were individually matched for age, body mass index, and waist/hip ratio with those who were normal glucose tolerant in a previous pregnancy (NGT). All women presented with normal glucose tolerance. Twenty pairs were submitted to the oral glucose tolerance test (OGTT) with plasma glucose, insulin, and C-peptide determinations. of the 20 pairs, 18 participated in hyperglycemic (10.0 mmol/l) clamp experiments with frequent plasma glucose and insulin determinations, allowing us to calculate first- and second-phase insulin release and the insulin sensitivity index. GDM and NGT women were compared using Student's t-test, the Mann-Whitney U-test, Friedman's non-parametric test, and the two proportion test for independent groups.Results. GDM women showed higher glycosylated hemoglobin values; at OGTT, they showed late insulin peak with increased plasma insulin levels only during the second hour, and a similar plasma C-peptide response despite a higher plasma glucose curve; during hyperglycemic clamp procedures, they showed similar biphasic insulin release and insulin sensitivity index. Considering that a woman with previous GDM had a defect in insulin release and/or insulin sensitivity, if its magnitude was at least 25% lower than that of the matched NGT woman, 43.8% showed impairment of first-phase insulin release and 55.6% insulin resistance.Conclusions. GDM women showed some degree of glucose intolerance. It is therefore necessary to follow them for a longer time.

Natural History of the Lutz's Frog Cycloramphus lutzorum Heyer, 1983 (Anura: Cycloramphidae) in the Brazilian Atlantic Forest: Description of the Advertisement Call, Tadpole, and Karyotype

We describe the advertisement call, tadpole, karyotype, and additional information on the natural history of Cycloramphus lutzorum from southern Brazil. Sonograms were generated from digitally recorded calls. Tadpoles were collected in the field for description in the lab, and an adult was collected for karyotyping. Data on seasonal activity were gathered monthly from November 2005 to November 2007. All tadpoles (N = 21), juveniles (N = 18), and adults (N = 52) were found exclusively in streams. Reproduction, as identified by calling frogs, occurred from July through November. Frogs call all day long, but mostly at dusk, from rock crevices inside the stream edges near the splash zone. The call is short and loud, with 11 pulsed notes, of 491-641 ms, with a dominant frequency of 0.98-1.39 kHz. We describe the exotrophic and semiterrestrial tadpoles, always found in constantly humid vertical rock walls in the stream. Tadpoles of C. lutzorum are recognized by differences in labial tooth row formula, eye diameter, body shape, position of nares, and development of tail. Like congeneric species, the karyotype of C. lutzorum comprises 26 metacentric and submetacentric chromosomes. Cycloramphus lutzorum is restricted to and adapted for living in fast flowing streams, many of which are threatened by deforestation, pollution, and habitat loss. Therefore, we recommend the status of C. lutzorum be changed from its current "Data Deficient" to "Near Threatened (NT)" in the IUCN species red list.